Title

Generating Yeast Two-Hybrid Bait Strains

UMMS Affiliation

Program in Systems Biology; Program in Molecular Medicine

Publication Date

2018-07-02

Document Type

Article

Disciplines

Biochemistry, Biophysics, and Structural Biology | Genetics and Genomics | Laboratory and Basic Science Research | Systems Biology

Abstract

Generating DNA-binding domain (DB)-bait strains for Gateway-compatible yeast two-hybrid (Y2H) screens involves three steps. The first is to generate an Entry clone containing a DNA fragment encoding the protein of interest (e.g., an open reading frame, ORF). The second is to transfer this DNA fragment from the Entry clone to the Y2H Destination vector, pDEST32. The final step is to transform this construct into the Y2H yeast strain, MaV103. This protocol takes 24-37 d plus sequence confirmation, if necessary, to complete.

DOI of Published Version

10.1101/pdb.prot094979

Source

Cold Spring Harb Protoc. 2018 Jul 2;2018(7):pdb.prot094979. doi: 10.1101/pdb.prot094979. Link to article on publisher's site

Journal/Book/Conference Title

Cold Spring Harbor protocols

Related Resources

Link to Article in PubMed

PubMed ID

29967272

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